Efficient Acquisition of Fully Human Antibody Genes against Self-Proteins by Sorting Single B Cells Stimulated with Vaccines Based on Nitrated T Helper Cell Epitopes

Figure 5

NitraTh improved peripheral human T cell activation and induces differentiation into Tfh cells in vivo. NCG mice were engrafted with PBMCs on day 0 and DCs on day 7 to build DC-HIS mice. Then, DC-HIS mice were immunized with HER2-NitraTh or HER2-Th on days 7 and 21. Splenocytes and lymph nodes were harvested from vaccinated mice on day 28. Data were expressed as (). (a, b, and e) FACS determined the percentages of multilineage human immune cells (pregated on hCD45⁺ cells as the total mononuclear cells) in the spleen. Helper T cells (gated as hCD4⁺), activated helper T cells (gated as hCD4⁺hCD69⁺), and B cells (gated as hCD19⁺). (d) FACS determined the percentages of Tfh cells (gated as CXCR5⁺ICOS⁺, pregated on hCD4⁺ cells) in the spleen. (c, f) Immunohistochemistry analyzed the distribution of hCD3⁺ T cells (brown) and hCD19⁺ B cells (brown) in the lymph node sections. Original magnification: ×400. n.s.: not significant. , , and compared with PBS; ^# and ^## compared with HER2-Th.

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